CN112807439A - Preparation method and application of implantable in-situ formed chitosan hydrogel - Google Patents

Preparation method and application of implantable in-situ formed chitosan hydrogel Download PDF

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CN112807439A
CN112807439A CN202110038527.3A CN202110038527A CN112807439A CN 112807439 A CN112807439 A CN 112807439A CN 202110038527 A CN202110038527 A CN 202110038527A CN 112807439 A CN112807439 A CN 112807439A
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chitosan
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acetic acid
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acid solution
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CN112807439B (en
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赵婧
王晓冉
丁涛
刘凯文
邓春昱
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Xian Polytechnic University
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Abstract

The invention discloses a preparation method of implantable in-situ formed chitosan hydrogel, which comprises the following steps: dissolving chitosan in an acetic acid solution to obtain an acetic acid solution of chitosan; preparing polyanion PNAM by NIPAm and AMPS free radical polymerization; under the condition of ice water bath, PNAM aqueous solution and NaHCO are mixed3Adding the solution into acetic acid solution of chitosan, stirring for 30min, placing the sample in a constant temperature water bath at 37 deg.C, gelatinizing, and freeze drying to obtain chitosan hydrogel. The chitosan is used as a carrier, has the advantages of bacteriostasis, sterilization, inflammation diminishing, hemostasis, wound healing promotion and the like, and the in-situ gel prepared by the polyelectrolyte composite method not only has temperature sensitivity, but also has the advantages ofHas pH sensitivity, can realize the controlled release of the drug, greatly reduces the trauma of the surgical operation and reduces the operation cost.

Description

Preparation method and application of implantable in-situ formed chitosan hydrogel
Technical Field
The invention belongs to the technical field of preparation of biomedical high polymer materials, and particularly relates to a preparation method of an implantable in-situ formed chitosan hydrogel, and an application of the chitosan hydrogel.
Background
Cardiovascular diseases are one of the major diseases seriously threatening the life and health of human beings, wherein the myocardial infarction patients have huge number and high death rate. Coronary artery interventional therapy, namely a large-wound operation such as a stent operation or a coronary artery bypass operation, is mostly adopted for treating myocardial infarction, and wound infection is easily caused after the operation, so that secondary injury is caused.
The implantable in-situ forming gel is one kind of tissue engineering scaffold, is in a solution flowing state at room temperature, can wrap therapeutic drugs, and is subjected to phase change at an injection part by virtue of environmental stimulation (light, temperature, pH value and the like) to form the gel after being injected to a designated position, so that the wound caused by a surgical operation is greatly reduced, the operation cost is reduced, and the implantable in-situ forming gel is a minimally invasive method convenient to operate and widely applied to the biomedical fields of drug delivery, tissue scaffold engineering and the like. However, the safety, retention in the body, etc. of the current carriers greatly limit the development of implantable materials.
The chitosan serving as a biocompatible and biodegradable cationic polysaccharide has the capabilities of sterilizing, inhibiting bacteria, diminishing inflammation, stopping bleeding and promoting wound healing, has biological adhesion and biological activity, can effectively increase the absorption of medicaments, improves the bioavailability of the medicaments, and is widely applied to the field of biomedicine. The novel implantable chitosan in-situ gel is constructed by taking chitosan as a carrier, has high safety compared with other solid preparations, is simple and feasible, does not need an operation and has small wound.
Disclosure of Invention
The invention aims to provide a preparation method of implantable in-situ forming chitosan hydrogel.
The invention also aims to provide the application of the chitosan hydrogel in controlled release of drugs.
The invention adopts the technical scheme that a preparation method of implantable in-situ forming chitosan hydrogel is implemented according to the following steps:
step 1, dissolving chitosan in an acetic acid solution to obtain an acetic acid solution of chitosan;
step 2, preparing polyanion PNAM by free radical polymerization of N-isopropylacrylamide (NIPAm) and 2-acrylamido-2-methylpropanesulfonic Acid (AMPS);
step 3, under the condition of ice water bath, mixing the aqueous solution of PNAM and NaHCO3Adding the solution into acetic acid solution of chitosan, stirring for 30min, placing the sample in a constant temperature water bath at 37 deg.C, gelatinizing, and freeze drying to obtain chitosan hydrogel.
The present invention is also characterized in that,
in the step 1, the mass fraction of the acetic acid solution is 1%; the concentration of the chitosan in the acetic acid solution is 15-30 mg/mL.
In the step 2, the method specifically comprises the following steps:
dissolving monomer NIPAm and monomer AMPS in mixed solvent, and adding initiator in N2Dropping the mixture into the mixed solvent under protection and stirring at a constant temperature of 60 ℃, wherein the dropping time is 2-3 h; and after the dropwise addition is finished, sealing the system for reaction for 24h, transferring the reaction solution into a dialysis bag with the molecular weight cutoff of 3500Da for dialysis for 3 days, and freeze-drying the sample at-46 ℃ to obtain the polyanion PNAM.
The initiator is azobisisoheptonitrile, and the mass of the initiator is 1 percent of the total mass of the monomer NIPAm and the monomer AMPS; the molar ratio of NIPAm to AMPS was 3: 7.
The volume ratio of the mixed solvent is 4: 1, distilled water and 1, 4-dioxane;
in step 3, NaHCO3The concentration of the solution is 0.4 mol/L; the mass concentration of the polyanionic PNAM aqueous solution is 3.0-6.0 mg/mL; acetic acid solution, PNAM solution and NaHCO of chitosan3The volume ratio of the solution is 10: 1-5: 5.
in the step 3, the temperature of the ice-water bath is 0-4 ℃, the gelation time is 60-500 s, the freeze drying time is 1-3 days, and the freeze drying temperature is-46 ℃.
The invention has the advantages that the chitosan is used as a carrier, the chitosan has the advantages of bacteriostasis, sterilization, inflammation diminishing, hemostasis, wound healing promotion and the like, the in-situ gel prepared by the polyelectrolyte complex method not only has temperature sensitivity, but also has pH sensitivity, the controlled release of the medicine can be realized, and the method is simple, feasible, nontoxic and safe. The implantable in-situ forming gel is in a solution flowing state at room temperature, and can form gel at a designated position when being injected into tissues or organs, thereby greatly reducing the trauma of surgical operation and lowering the operation cost.
Drawings
FIG. 1 is a schematic diagram of the formation process of the implantable in situ formed chitosan hydrogel of the present invention;
FIG. 2 is an infrared spectrum of a chitosan hydrogel prepared by the polyelectrolyte complex method of the present invention;
FIG. 3 is an SEM image of a chitosan hydrogel prepared by the polyelectrolyte complexing method according to the present invention;
FIG. 4 is an inverted microscope photograph of the chitosan hydrogel of the present invention cultured with cells for 24 h;
FIG. 5 is an inverted micrograph of positive control phenol after 24h incubation with cells;
fig. 6 is a drug release profile of aspirin-loaded chitosan hydrogel.
Detailed Description
The present invention will be described in detail below with reference to the accompanying drawings and specific embodiments.
The invention relates to a preparation method of implantable in-situ formed chitosan hydrogel, which is implemented according to the following steps:
step 1, dissolving chitosan in an acetic acid solution to obtain an acetic acid solution of chitosan;
the mass fraction of the acetic acid solution is 1 percent;
the concentration of the chitosan in the acetic acid solution is 15-30 mg/mL; the viscosity of the chitosan is medium-viscosity chitosan (200-400 mPa.s) or high-viscosity chitosan (>400 mPa.s);
step 2, preparing polyanion PNAM by free radical polymerization of N-isopropylacrylamide (NIPAm) and 2-acrylamido-2-methylpropanesulfonic Acid (AMPS), which comprises the following steps:
dissolving NIPAm monomer and AMPS monomer in mixed solvent, mixing initiator and monomer mixture, and adding N solution2Protective, stirring at a constant temperature of 60 ℃, dropwise adding into a three-neck flask, sealing the system to react for 24 hours after dropwise adding, transferring the reaction liquid into a dialysis bag with a molecular weight cutoff of 3500Da for dialysis for 3 days, and freeze-drying the sample at-46 ℃ to obtain polyanion PNAM;
the dripping time is 2-3 h; the initiator is Azobisisoheptonitrile (AVBN), and the mass of the initiator is 1 percent of the total mass of the monomer NIPAm and the monomer AMPS;
the volume ratio of the mixed solvent is 4: 1, distilled water and 1, 4-dioxane;
wherein, the molar ratio of NIPAm to AMPS is 3:7, and the total concentration of the monomers in the mixed solvent is 0.5 mol/L;
step 3, under the condition of ice water bath, mixing the aqueous solution of PNAM and NaHCO3Adding the solution into acetic acid solution of chitosan, stirring for 30min, placing the sample in a constant-temperature water bath kettle at 37 deg.C, gelatinizing, and freeze drying to obtain chitosan hydrogel;
NaHCO3the concentration of the solution is 0.4 mol/L; the mass concentration of the polyanionic PNAM aqueous solution is 3.0-6.0 mg/mL;
the temperature of the ice-water bath is 0-4 ℃, the gelation time is 60-500 s, the freeze drying time is 1-3 days, and the freeze drying temperature is-46 ℃;
acetic acid solution, PNAM solution and NaHCO of chitosan3The volume ratio of the solution is 10: 1-5: 5.
the method of the invention uses chitosan as a carrier, and adopts a polyelectrolyte complex method to construct implantable chitosan in-situ gel through electrostatic interaction between the chitosan and a functional polymer containing a temperature-sensitive monomer N-isopropyl acrylamide (NIPAm), thereby realizing site-specific injection and site-specific release.
Fig. 1 is a schematic diagram of a forming process of an implantable in-situ forming chitosan hydrogel, wherein the critical temperature is 35-37 ℃, the chitosan hydrogel is in an injectable flowing solution state below the critical temperature, and gel is formed within a certain time after the critical temperature is reached.
FIG. 2 is an infrared spectrum of 3440cm for chitosan hydrogel prepared by polyelectrolyte complex method-1Has a reduced absorption peak of 1601cm-1The amino group shock absorption peak disappeared, and 1650cm-1The vibration absorption peak of the carbonyl group on the amide group is 1400cm-1Is the C-N stretching vibration absorption peak; 1157cm-1、1074cm-1Stretching vibration of S ═ O in the sulfonic acid group, 514cm-1And a C-S stretching vibration absorption peak appears. Shows that the interaction between the amino group on the chitosan molecular chain and the polymer occurs, and successfully prepares the polymer crosslinked chitosan hydrogel.
FIG. 3 is an SEM image of chitosan hydrogel prepared by polyelectrolyte complex method, the surface of which is layered and porous, and has a cross-linked network and a multi-layer honeycomb structure. The layered structure can be applied to a stent material, and the porous shape is more suitable for drug loading, adsorption and other properties.
Fig. 4 and 5 are inverted microscope photographs of L929 cells as cell models, after culturing positive control phenol and cells for 24h by using MTT method to evaluate chitosan hydrogel, the positive control phenol group cells are mostly dead, the chitosan hydrogel group cells are distributed closely, are mostly fusiform, and are full, further explaining that the chitosan hydrogel group cells have good biocompatibility, and simultaneously, the cells are found to play a role in promoting cell proliferation.
The chitosan hydrogel is applied to controlled release of drugs. Weighing chitosan, dissolving in 1% acetic acid solution (20mg/mL), standing for removing bubbles after complete dissolution, cooling to 4 ℃ under ice bath condition, dissolving 0.01g aspirin (ASA) in a mixed solution of 4.0mg/mL PNAM aqueous solution and 0.4M sodium bicarbonate, dropwise adding into an injector in an ice water bath while stirring until the pH is about 7.2, and stirring for reaction for 30 min. The mixed solution was injected into a test tube having a diameter of 10mm in an amount of 2mL by a syringe, and placed in a constant temperature water bath at 37 ℃ for a gelation time of 150 seconds. The drug loading rate of the chitosan hydrogel is 12.6 percent, and the encapsulation rate is 62.8 percent.
A certain amount of drug-loaded hydrogel is placed in a centrifuge tube of 25mL of PBS buffer solution with pH values of 4, 5.5 and 7.4 and fixed in a constant temperature shaking table, and the temperature and the rotating speed of the shaking table are respectively set to be 37 ℃ and 100 rpm. 5mL of the release solution was removed at intervals, and 5mL of PBS buffer was added. The release liquid is tested by adopting an ultraviolet-visible spectrometer, and a release rate curve under different pH conditions is obtained by calculation, so that slow release and controllable release are realized under certain conditions, and the curative effect of the medicament is improved.
Fig. 6 is a drug release curve of aspirin-loaded chitosan hydrogel, and the influence curves of buffer solutions with different release mediums (pH 4, pH 5.5, pH 7.4) on the controlled release performance of aspirin at 37 ℃. The slow release of the aspirin is realized after 2h, the release rate of the aspirin is fastest under the condition that the pH is 4, and the release of the aspirin can be controlled according to different release rates under different pH conditions.
Example 1
The invention relates to a preparation method of implantable in-situ formed chitosan hydrogel, which is implemented according to the following steps:
step 1, dissolving chitosan in an acetic acid solution to obtain an acetic acid solution of chitosan;
the mass fraction of the acetic acid solution is 1 percent; the concentration of the chitosan in the acetic acid solution is 20 mg/mL;
step 2, preparing polyanion PNAM by free radical polymerization of N-isopropylacrylamide (NIPAm) and 2-acrylamido-2-methylpropanesulfonic Acid (AMPS), which comprises the following steps:
dissolving monomer NIPAm and monomer AMPS in mixed solvent, and adding initiator in N2Protective, stirring at a constant temperature of 60 ℃, dropwise adding into the mixed solvent, sealing the system to react for 24h after dropwise adding, transferring the reaction solution into a dialysis bag with a molecular weight cutoff of 3500Da for dialysis for 3 days, and freeze-drying the sample at-46 ℃ to obtain polyanion PNAM;
the dripping time is 2 hours; the initiator is Azobisisoheptonitrile (AVBN), and the mass of the initiator is 1 percent of the total mass of the monomer NIPAm and the monomer AMPS;
the volume ratio of the mixed solvent is 4: 1, distilled water and 1, 4-dioxane;
wherein, the molar ratio of NIPAm to AMPS is 3:7, and the total concentration of the monomers in the mixed solvent is 0.5 mol/L;
step 3, under the condition of ice water bath, mixing the aqueous solution of PNAM and NaHCO3Adding the solution into acetic acid solution of chitosan, stirring for 30min, placing the sample in a constant-temperature water bath kettle at 37 deg.C, gelatinizing, and freeze drying to obtain chitosan hydrogel;
NaHCO3the concentration of the solution is 0.4 mol/L; the mass concentration of the polyanionic PNAM aqueous solution is 3.0 mg/mL;
the temperature of the ice-water bath is 0 ℃, the gelation time is 180s, the freeze drying time is 1 day, and the freeze drying temperature is-46 ℃;
acetic acid solution, PNAM solution and NaHCO of chitosan3The volume ratio of the solution is 10: 2: 5.
example 2
The invention relates to a preparation method of implantable in-situ formed chitosan hydrogel, which is implemented according to the following steps:
step 1, dissolving chitosan in an acetic acid solution to obtain an acetic acid solution of chitosan;
the mass fraction of the acetic acid solution is 1 percent;
the concentration of the chitosan in the acetic acid solution is 15 mg/mL; the viscosity of the chitosan is medium-viscosity chitosan;
step 2, preparing polyanion PNAM by free radical polymerization of N-isopropylacrylamide (NIPAm) and 2-acrylamido-2-methylpropanesulfonic Acid (AMPS), which comprises the following steps:
dissolving monomer NIPAm and monomer AMPS in mixed solvent, and adding initiator in N2Protective, stirring at a constant temperature of 60 ℃, dropwise adding into the mixed solvent, sealing the system to react for 24h after dropwise adding, transferring the reaction solution into a dialysis bag with a molecular weight cutoff of 3500Da for dialysis for 3 days, and freeze-drying the sample at-46 ℃ to obtain polyanion PNAM;
the dripping time is 2 hours; the initiator is Azobisisoheptonitrile (AVBN), and the mass of the initiator is 1 percent of the total mass of the monomer NIPAm and the monomer AMPS;
the volume ratio of the mixed solvent is 4: 1, distilled water and 1, 4-dioxane;
wherein, the molar ratio of NIPAm to AMPS is 3:7, and the total concentration of the monomers in the mixed solvent is 0.5 mol/L;
step 3, under the condition of ice water bath, mixing the aqueous solution of PNAM and NaHCO3Adding the solution into acetic acid solution of chitosan, stirring for 30min, placing the sample in a constant-temperature water bath kettle at 37 deg.C, gelatinizing, and freeze drying to obtain chitosan hydrogel;
NaHCO3the concentration of the solution is 0.4 mol/L; the mass concentration of the polyanionic PNAM aqueous solution is 3.0 mg/mL;
the temperature of the ice-water bath is 1 ℃, the gelation time is 340s, the freeze drying time is 2 days, and the freeze drying temperature is-46 ℃;
acetic acid solution, PNAM solution and NaHCO of chitosan3The volume ratio of the solution is 10: 2: 5;
example 3
The invention relates to a preparation method of implantable in-situ formed chitosan hydrogel, which is implemented according to the following steps:
step 1, dissolving chitosan in an acetic acid solution to obtain an acetic acid solution of chitosan;
the mass fraction of the acetic acid solution is 1 percent;
the concentration of the chitosan in the acetic acid solution is 20 mg/mL; the chitosan is high viscosity chitosan (>400 mpa.s);
step 2, preparing polyanion PNAM by free radical polymerization of N-isopropylacrylamide (NIPAm) and 2-acrylamido-2-methylpropanesulfonic Acid (AMPS), which comprises the following steps:
dissolving monomer NIPAm and monomer AMPS in mixed solvent, and adding initiator in N2Protective, stirring at a constant temperature of 60 ℃, dropwise adding into the mixed solvent, sealing the system to react for 24h after dropwise adding, transferring the reaction solution into a dialysis bag with a molecular weight cutoff of 3500Da for dialysis for 3 days, and freeze-drying the sample at-46 ℃ to obtain polyanion PNAM;
the dripping time is 2 hours; the initiator is Azobisisoheptonitrile (AVBN), and the mass of the initiator is 1 percent of the total mass of the monomer NIPAm and the monomer AMPS;
the volume ratio of the mixed solvent is 4: 1, distilled water and 1, 4-dioxane;
wherein, the molar ratio of NIPAm to AMPS is 3:7, and the total concentration of the monomers in the mixed solvent is 0.5 mol/L;
step 3, under the condition of ice water bath, mixing the aqueous solution of PNAM and NaHCO3Adding the solution into acetic acid solution of chitosan, stirring for 30min, placing the sample in a constant-temperature water bath kettle at 37 deg.C, gelatinizing, and freeze drying to obtain chitosan hydrogel;
NaHCO3the concentration of the solution is 0.4 mol/L; the mass concentration of the polyanionic PNAM aqueous solution is 4.0 mg/mL;
the temperature of the ice-water bath is 1 ℃, the gelation time is 120s, the freeze drying time is 1 day, and the freeze drying temperature is-46 ℃;
acetic acid solution, PNAM solution and NaHCO of chitosan3The volume ratio of the solution is 10: 3: 5;
example 4
The invention relates to a preparation method of implantable in-situ formed chitosan hydrogel, which is implemented according to the following steps:
step 1, dissolving chitosan in an acetic acid solution to obtain an acetic acid solution of chitosan;
the mass fraction of the acetic acid solution is 1 percent;
the concentration of the chitosan in the acetic acid solution is 25 mg/mL; the chitosan is medium viscosity chitosan;
step 2, preparing polyanion PNAM by free radical polymerization of N-isopropylacrylamide (NIPAm) and 2-acrylamido-2-methylpropanesulfonic Acid (AMPS), which comprises the following steps:
dissolving monomer NIPAm and monomer AMPS in mixed solvent, and adding initiator in N2Protective, stirring at a constant temperature of 60 ℃, dropwise adding into the mixed solvent, sealing the system to react for 24h after dropwise adding, transferring the reaction solution into a dialysis bag with a molecular weight cutoff of 3500Da for dialysis for 3 days, and freeze-drying the sample at-46 ℃ to obtain polyanion PNAM;
the dripping time is 2.5 h; the initiator is Azobisisoheptonitrile (AVBN), and the mass of the initiator is 1 percent of the total mass of the monomer NIPAm and the monomer AMPS;
the volume ratio of the mixed solvent is 4: 1, distilled water and 1, 4-dioxane;
wherein, the molar ratio of NIPAm to AMPS is 3:7, and the total concentration of the monomers in the mixed solvent is 0.5 mol/L;
step 3, under the condition of ice water bath, mixing the aqueous solution of PNAM and NaHCO3Adding the solution into acetic acid solution of chitosan, stirring for 30min, placing the sample in a constant-temperature water bath kettle at 37 deg.C, gelatinizing, and freeze drying to obtain chitosan hydrogel;
NaHCO3the concentration of the solution is 0.4 mol/L; the mass concentration of the polyanionic PNAM aqueous solution is 5.0 mg/mL;
the temperature of the ice-water bath is 2 ℃, the gelation time is 80s, the freeze drying time is 1 day, and the freeze drying temperature is-46 ℃;
acetic acid solution, PNAM solution and NaHCO of chitosan3The volume ratio of the solution is 10: 3: 5;
example 5
The invention relates to a preparation method of implantable in-situ formed chitosan hydrogel, which is implemented according to the following steps:
step 1, dissolving chitosan in an acetic acid solution to obtain an acetic acid solution of chitosan;
the mass fraction of the acetic acid solution is 1 percent;
the concentration of the chitosan in the acetic acid solution is 15 mg/mL; the chitosan is high viscosity chitosan (>400 mpa.s);
step 2, preparing polyanion PNAM by free radical polymerization of N-isopropylacrylamide (NIPAm) and 2-acrylamido-2-methylpropanesulfonic Acid (AMPS), which comprises the following steps:
dissolving monomer NIPAm and monomer AMPS in mixed solvent, and adding initiator in N2Protective, stirring at a constant temperature of 60 ℃, dropwise adding into the mixed solvent, sealing the system to react for 24h after dropwise adding, transferring the reaction solution into a dialysis bag with a molecular weight cutoff of 3500Da for dialysis for 3 days, and freeze-drying the sample at-46 ℃ to obtain polyanion PNAM;
the dripping time is 3 h; the initiator is Azobisisoheptonitrile (AVBN), and the mass of the initiator is 1 percent of the total mass of the monomer NIPAm and the monomer AMPS;
the volume ratio of the mixed solvent is 4: 1, distilled water and 1, 4-dioxane;
wherein, the molar ratio of NIPAm to AMPS is 3:7, and the total concentration of the monomers in the mixed solvent is 0.5 mol/L;
step 3, under the condition of ice water bath, mixing the aqueous solution of PNAM and NaHCO3Adding the solution into acetic acid solution of chitosan, stirring for 30min, placing the sample in a constant-temperature water bath kettle at 37 deg.C, gelatinizing, and freeze drying to obtain chitosan hydrogel;
NaHCO3the concentration of the solution is 0.4 mol/L; the mass concentration of the polyanionic PNAM aqueous solution is 6.0 mg/mL;
the temperature of the ice-water bath is 4 ℃, the gelation time is 450s, the freeze drying time is 3 days, and the freeze drying temperature is-46 ℃;
acetic acid solution, PNAM solution and NaHCO of chitosan3The volume ratio of the solution is 10: 4: 5.

Claims (8)

1. a preparation method of implantable in-situ forming chitosan hydrogel is characterized by comprising the following steps:
step 1, dissolving chitosan in an acetic acid solution to obtain an acetic acid solution of chitosan;
step 2, preparing polyanion PNAM by polymerizing N-isopropyl acrylamide NIPAm and 2-acrylamide-2-methylpropanesulfonic acid AMPS free radicals;
step 3, under the condition of ice water bath, mixing the aqueous solution of PNAM and NaHCO3Adding the solution into acetic acid solution of chitosan, stirring for 30min, placing the sample in a constant temperature water bath at 37 deg.C, gelatinizing, and freeze drying to obtain chitosan hydrogel.
2. The method for preparing the implantable in-situ forming chitosan hydrogel as claimed in claim 1, wherein in the step 1, the mass fraction of the acetic acid solution is 1%; the concentration of the chitosan in the acetic acid solution is 15-30 mg/mL.
3. The method for preparing the implantable in-situ formed chitosan hydrogel according to claim 1, wherein the step 2 specifically comprises:
dissolving NIPAm monomer and AMPS monomer in mixed solvent, mixing initiator and monomer mixture, and adding N solution2Dropping the mixture into a three-neck flask under protection and stirring at a constant temperature of 60 ℃, wherein the dropping time is 2-3 h; and after the dropwise addition is finished, sealing the system for reaction for 24h, transferring the reaction solution into a dialysis bag with the molecular weight cutoff of 3500Da for dialysis for 3 days, and freeze-drying the sample at-46 ℃ to obtain the polyanion PNAM.
4. The method for preparing the implantable in-situ forming chitosan hydrogel as claimed in claim 3, wherein the initiator is azobisisoheptonitrile, and the mass of the initiator is 1% of the total mass of the monomer NIPAm and the monomer AMPS; the molar ratio of NIPAm to AMPS was 3: 7.
5. The method for preparing the implantable in-situ forming chitosan hydrogel according to claim 3, wherein the mixed solvent is prepared from the following components in a volume ratio of 4: 1 distilled water and 1, 4-dioxane.
6. The method for preparing implantable in-situ forming chitosan hydrogel as claimed in claim 1, wherein in the step 3, NaHCO is used3The concentration of the solution is 0.4 mol/L; the mass concentration of the polyanionic PNAM aqueous solution is 3.0-6.0 mg/mL; acetic acid solution, PNAM solution and NaHCO of chitosan3The volume ratio of the solution is 10: 1-5: 5.
7. the method for preparing the implantable in-situ forming chitosan hydrogel according to claim 1, wherein in the step 3, the temperature of the ice-water bath is 0-4 ℃, the gelation time is 60-500 s, the freeze-drying time is 1-3 days, and the freeze-drying temperature is-46 ℃.
8. An implantable in situ forming chitosan hydrogel according to claim 1, wherein the chitosan hydrogel is used for controlled release of a drug.
CN202110038527.3A 2021-01-12 2021-01-12 Preparation method and application of implantable in-situ formed chitosan hydrogel Active CN112807439B (en)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN116328022A (en) * 2023-02-27 2023-06-27 西安工程大学 Preparation method and application of injectable mussel-like chitosan hydrogel adhesive
CN117646329A (en) * 2024-01-29 2024-03-05 比音勒芬服饰股份有限公司 Intelligent heat and humidity regulating fiber and fabric comprising same

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
JING ZHAO ET AL: "Dual ionically crosslinked chitosan-based injectable hydrogel as drug delivery system", 《COLLOID AND POLYMER SCIENCE》 *
赵婧 等: "温度/pH双重响应型壳聚糖纳米药物载体的制备及性能", 《功能材料》 *

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN116328022A (en) * 2023-02-27 2023-06-27 西安工程大学 Preparation method and application of injectable mussel-like chitosan hydrogel adhesive
CN117646329A (en) * 2024-01-29 2024-03-05 比音勒芬服饰股份有限公司 Intelligent heat and humidity regulating fiber and fabric comprising same
CN117646329B (en) * 2024-01-29 2024-03-29 比音勒芬服饰股份有限公司 Intelligent heat and humidity regulating fiber and fabric comprising same

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